RESEARCH II/Sci & Tech

CMSHS

LAB EXPERIMENT
Animal Research

PRACTICAL RODENT INJECTION TECHNIQUES

GUIDELINES
The procedures described in this SOP should be followed when liquids will be administered to mice, guinea
pigs, or hamsters by intraperitoneal and subcutaneous. All substances must be sterile and administered using sterile
instruments to minimize the risk of infection. Maximum injection volumes and needle sizes must be followed to
minimize trauma to tissues and discomfort for the animals.
Maximum injection volumes
Mice
Route

Volume

Guinea Pigs
Needle size

(mL)

Volume

Needle size

(mL)

Hamsters
Volume

Needle size

(mL)

Intraperitoneal (IP)

2.0

25G

10ml/kg

23G

3-4

23G

Subcutaneous (SC)

2.0

25G

5ml/kg

23G

3-4

23G

MATERIALS

SUBSTANCES/FLUIDS

Sterile needles- 23G, 25G, 27G

Saline Solution (1tsp 100 mL)

Sterile syringes- CC, 1CC, 3CC, 5CC, 10CC

Clippers
70% ethanol
Gauze
PRECAUTIONS

Always wear your PPEs

Rodents have the tendency to defend itself so please wear proper gloves.

Dispose the used needles properly. DO NOT RE-USE IT

Procedures in the mouse can generally be performed by one person. For awake procedures in rats and hamsters
it may be easier to have two people, one to restrain the animal and one to administer the injection.
Perform the experiments only after the animals stabilize. Let them rest for a period of 1 day. Hence, each
procedure should be performed one at a time

PROCEDURE
Dissolve the saline solution in a 100 mL beaker. Make sure that the solution will have the right
consistency before administering it to the specimen. While waiting for it to dissolve, prepare for the next
procedure.

Intraperitoneal injection:
1. Prepare the syringe and needle with the inoculum, removing the cap from the needle and expelling any air
bubbles.
2. Manually restrain the animal by grasping the lose skin at the scruff of the neck with the thumb and forefinger
to immobilize the head and torso. Pick the animal up and turn it so the abdomen is facing up, supporting the
back and hind end.
3. Wipe the lower left quadrant of the abdomen with 70% alcohol and allow to dry.
4. Tip the animal so the nose is pointing down slightly then insert the needle at a 30 angle in the lower left
quadrant of the abdomen.
5. Retract the plunger slightly and if any fluid is aspirated remove the needle and prepare a new inoculum before
repeating the injection.
6. When the needle is correctly positioned, slowly inject the inoculum then remove the needle.
7. Once the injection is complete, release the animal into the cage.
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RESEARCH II/Sci & Tech

CMSHS

LAB EXPERIMENT
Animal Research

Subcutaneous injection
1. Prepare the syringe and needle with the inoculum, removing the cap from the needle and expelling any air
bubbles.
2. Manually restrain the animal by grasping the lose skin at the scruff of the neck with the thumb and forefinger
to immobilize the head and torso. Place the animal on a flat surface to stabilize the body.
3. Gently pull up the skin over the desired injection site, i.e. nape of neck, flank, or rump, to create a small pocket.
4. Insert the needle into the pocket and retract the plunger slightly. If any fluid or blood is seen, remove the needle
and reposition it.
5. Depress the plunger slightly and if any resistance is felt remove the needle and reposition it.
6. Once the needle is correctly positioned, slowly inject the inoculum then remove the needle.
7. Once the injection is complete release the animal into the cage.
POST PROCEDURAL MONITORING
1. Anesthetized animals must be monitored continuously until they are awake and able to walk normally.
2. Awake animals should be monitored at a minimum 1 hour and 24 hours after the procedure .
3. Observe the animals for the following signs:
a. Bleeding from the injection site
b. Labored breathing
c. Discharge from the nose or mouth
d. Ruffled fur
e. Hunched posture
f.

Lethargy

g. Minimal responsiveness
4. Lethargy, labored breathing, minimal responsiveness and uncontrolled bleeding or discharge are criteria for
immediate euthanasia.
5. Document all observations on the experimental health monitoring sheet. Alert the Attending Veterinarian of
any ill or injured mice.
ADVANCE STUDY TASKS
1. What is intraperitoneal and subcutaneous injection?

2. Draw the set-up for both intraperitoneal and subcutaneous injection

3. How would improper injection volumes and needle sizes result trauma to tissues and discomfort to the animals?

Checked by: ________________________

salongaeugene@gmail.com

RESEARCH II/Sci & Tech

CMSHS

LAB EXPERIMENT
Animal Research

LABORATORY REPORT SHEET

Group Code: ________ Grade & Section: _________________
Leader: ____________________________ Code: ___________
Name: ____________________________ Code: ___________
Name: ____________________________ Code: ___________
Name: ____________________________ Code: ___________

Criteria:
TOTAL _____________
Penmanship (20%) ___________
Grammar (20%) ___________
Presentation (20%).... ___________
Content (40%)... ___________

DATA AND RESULTS

Qualitative Data:
Describe the Animals as the solution are injected:
Mice ______________________________________________________________________________________
Hamster___________________________________________________________________________________________
Guinea Pig _______________________________________________________________________________________

Quantitative Data:
Table 1: Post Procedural Monitoring Table
Animal

Procedure

Behavioral Observations
5 minutes

1 hour

3 hours

24 hours

Intraperitoneal
Mice
Subcutaneous
Intraperitoneal
Guinea Pig
Subcutaneous
Intraperitoneal
Hamster
Subcutaneous
SUMMARY OF FINDINGS

GUIDE QUESTIONS
1. Why is it important to sterilize the needles before experimentation?

2. For intraperitoneal injection, why should the needle be injected at a 30 angle in the lower left quadrant of the
abdomen of the specimen?

3. Why should the needle be removed and repositioned when a resistance is felt?

CONCLUSIONS AND RECOMMENDATIONS:

REFERENCES:

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salongaeugene@gmail.com